Connection between protein-tyrosine kinase inhibition and coping with oxidative stress in Bacillus subtilis.

In bacteria, attenuation of protein-tyrosine phosphorylation occurs during oxidative stress. The main described mechanism behind this effect is the H2O2-triggered conversion of bacterial phospho-tyrosines to protein-bound 3,4-dihydroxyphenylalanine. This disrupts the bacterial tyrosine phosphorylation-based signaling network, which alters the bacterial polysaccharide biosynthesis. Herein, we report an alternative mechanism, in which oxidative stress leads to a direct inhibition of bacterial protein-tyrosine kinases (BY-kinases). We show that DefA, a minor peptide deformylase, inhibits the activity of BY-kinase PtkA when Bacillus subtilis is exposed to oxidative stress. High levels of PtkA activity are known to destabilize B. subtilis pellicle formation, which leads to higher sensitivity to oxidative stress. Interaction with DefA inhibits both PtkA autophosphorylation and phosphorylation of its substrate Ugd, which is involved in exopolysaccharide formation. Inactivation of defA drastically reduces the capacity of B. subtilis to cope with oxidative stress, but it does not affect the major oxidative stress regulons PerR, OhrR, and Spx, indicating that PtkA inhibition is the main pathway for DefA involvement in this stress response. Structural analysis identified DefA residues Asn95, Tyr150, and Glu152 as essential for interaction with PtkA. Inhibition of PtkA depends also on the presence of a C-terminal α-helix of DefA, which resembles PtkA-interacting motifs from known PtkA activators, TkmA, SalA, and MinD. Loss of either the key interacting residues or the inhibitory helix of DefA abolishes inhibition of PtkA in vitro and impairs postoxidative stress recovery in vivo, confirming the involvement of these structural features in the proposed mechanism.

Antibacterial applications of biologically synthesized Pichia pastoris silver nanoparticles.

Objectives: This article highlights the biological synthesis of silver nanoparticles (AgNPs) with their characteristic analysis, and it focuses on the application of synthesized NPs against multidrug resistance (MDR) bacteria. A cytotoxicity study was performed to assess the biocompatibility. Methods: Silver nanoparticle (AgNPs) formation was confirmed by different characterization methods such as UV-Vis spectrophotometer, Dynamic light scattering (DLS)- Zeta, Fourier transform infrared (FTIR), and Transmission electron microscope (TEM). The antimicrobial activity of the AgNPs was checked against various bacterial strains of Staphylococcus aureus (S. aureus), Escherichia coli (E. coli), Enterococcus faecalis (E. faecalis), and Klebsiella pneumonia (K. pneumonia) by disc diffusion, minimum inhibition concentration test (MIC), and kinetic studies. The cytotoxicity of NPs against the Vero cell line was studied by cytotoxic assay. Results: The primary analysis of the formation of nanoparticles (NPs) was made by UV-Vis spectrophotometric analysis at 400 nm. At the same time, the efficient capping checked by FTIR shows the presence of a functional group at different wavelengths 3284, 1641,1573,1388,1288, and 1068 cm-1. At the same time, the transmission electron microscopic analysis (TEM) and DLS show that the shape and size of the synthesized NPs possess an average size of around ∼10-30 nm with spherical morphology. Further, the zeta potential confirmed the stability of the NPs. While the yield of NPs formation from silver salt was determined by an online yield calculator with the EDX analysis results. Synthesized NPs showed bactericidal effects against all the selected MDR pathogens with nontoxic effects against mammalian cells. Conclusion: Our findings indicate the remarkable antimicrobial activity of the biologically synthesized AgNPs, which can be an antimicrobial agent against multi-drug-resistant bacteria.

Differences in interaction of graphene/graphene oxide with bacterial and mammalian cell membranes.

Graphene, a single layer, hexagonally packed two-dimensional carbon sheet is an attractive candidate for diverse applications including antibacterial potential and drug delivery. One of the knowledge gaps in biomedical application of graphene is the interaction of these materials with the cells. To address this, we investigated the interaction between graphene materials (graphene and graphene oxide) and plasma membranes of cells (bacterial and mammalian cells). The interactions of four of the most abundant phospholipids in bacteria and mammalian plasma membranes with graphene materials were studied using density functional theory (DFT) at the atomic level. The calculations showed that the mammalian phospholipids have stronger bonding to each other compared to bacterial phospholipids. When the graphene/graphene oxide sheet is approaching the phospholipid pairs, the bacterial pairs exhibit less repulsive interactions, thereby a more stable system with the sheets was found. We also assembled bacterial and mammalian phospholipids into liposomes. We further observed that the bacterial liposomes and cells let the graphene flakes penetrate the membrane. The differential scanning calorimetry measurements of liposomes revealed that the bacterial liposomes have the lowest heat capacity; this strengthens the theoretical predictions of weaker interaction between the bacterial phospholipids compared to the mammalian phospholipids. We further demonstrated that graphene oxide could be internalized into the mammalian liposomes without disrupting the membrane integrity. The results suggest that the weak bonding among bacteria phospholipids and less repulsive force when graphene materials approach, result in graphene materials interacting differently with the bacteria compared to mammalian cells.

Biomimetic Antibacterial Gelatin Hydrogels with Multifunctional Properties for Biomedical Applications.

A facile novel approach of introducing dopamine and [2-(methacryloyloxy) ethyl] dimethyl-(3-sulfopropyl) ammonium hydroxide via dopamine-triggered in situ synthesis into gelatin hydrogels in the presence of ZnSO4 is presented in this study. Remarkably, the resulting hydrogels showed 99.99 and 100% antibacterial efficiency against Gram-positive and Gram-negative bacteria, respectively, making them the highest performing surfaces in their class. Furthermore, the hydrogels showed adhesive properties, self-healing ability, antifreeze properties, electrical conductivity, fatigue resistance, and mechanical stability from -100 to 80 °C. The added multifunctional performance overcomes several disadvantages of gelatin-based hydrogels such as poor mechanical properties and limited thermostability. Overall, the newly developed hydrogels show significant potential for numerous biomedical applications, such as wearable monitoring sensors and antibacterial coatings.

Polysaccharide-based antibacterial coating technologies.

To tackle antimicrobial resistance, a global threat identified by the United Nations, is a common cause of healthcare-associated infections (HAI) and is responsible for significant costs on healthcare systems, a substantial amount of research has been devoted to developing polysaccharide-based strategies that prevent bacterial attachment and biofilm formation on surfaces. Polysaccharides are essential building blocks for life and an abundant renewable resource that have attracted much attention due to their intrinsic remarkable biological potential antibacterial activities. If converted into efficient antibacterial coatings that could be applied to a broad range of surfaces and applications, polysaccharide-based coatings could have a significant potential global impact. However, the ultimate success of polysaccharide-based antibacterial materials will be determined by their potential for use in manufacturing processes that are scalable, versatile, and affordable. Therefore, in this review we focus on recent advances in polysaccharide-based antibacterial coatings from the perspective of fabrication methods. We first provide an overview of strategies for designing polysaccharide-based antimicrobial formulations and methods to assess the antibacterial properties of coatings. Recent advances on manufacturing polysaccharide-based coatings using some of the most common polysaccharides and fabrication methods are then detailed, followed by a critical comparative overview of associated challenges and opportunities for future developments. STATEMENT OF SIGNIFICANCE: Our review presents a timely perspective by being the first review in the field to focus on advances on polysaccharide-based antibacterial coatings from the perspective of fabrication methods along with an overview of strategies for designing polysaccharide-based antimicrobial formulations, methods to assess the antibacterial properties of coatings as well as a critical comparative overview of associated challenges and opportunities for future developments. Meanwhile this work is specifically targeted at an audience focused on featuring critical information and guidelines for developing polysaccharide-based coatings. Including such a complementary work in the journal could lead to further developments on polysaccharide antibacterial applications.

Antibiotic-Loaded Boron Nitride Nanoconjugate with Strong Performance against Planktonic Bacteria and Biofilms.

Protecting surfaces from biofilm formation presents a significant challenge in the biomedical field. The utilization of antimicrobial component-conjugated nanoparticles is becoming an attractive strategy against infectious biofilms. Boron nitride (BN) nanomaterials have a unique biomedical application value due to their excellent biocompatibility. Here, we developed antibiotic-loaded BN nanoconjugates to combat bacterial biofilms. Antibiofilm testing included two types of pathogens, Staphylococcus aureus and Escherichia coli. Gentamicin was loaded on polydopamine-modified BN nanoparticles (GPBN) to construct a nanoconjugate, which was very effective in killing E. coli and S. aureus planktonic cells. GPBN exhibited equally strong capacity for biofilm destruction, tested on preformed biofilms. A 24 h treatment with the nanoconjugate reduced cell viability by more than 90%. Our results suggest that GPBN adheres to the surface of the biofilm, penetrates inside the biofilm matrix, and finally deactivates the cells. Interestingly, the GPBN coatings also strongly inhibited the formation of bacterial biofilms. Based on these results, we suggest that GPBN could serve as an effective means for treating biofilm-associated infections and as coatings for biofilm prevention.

Polymyxin B complexation enhances the antimicrobial potential of graphene oxide.

Introduction: The antibacterial activity of graphene oxide (GO) has been widely explored and tested against various pathogenic bacterial strains. Although antimicrobial activity of GO against planktonic bacterial cells was demonstrated, its bacteriostatic and bactericidal effect alone is not sufficient to damage sedentary and well protected bacterial cells inside biofilms. Thus, to be utilized as an effective antibacterial agent, it is necessary to improve the antibacterial activity of GO either by integration with other nanomaterials or by attachment of antimicrobial agents. In this study, antimicrobial peptide polymyxin B (PMB) was adsorbed onto the surface of pristine GO and GO functionalized with triethylene glycol. Methods: The antibacterial effects of the resulting materials were examined by evaluating minimum inhibitory concentration, minimum bactericidal concentration, time kill assay, live/dead viability staining and scanning electron microscopy. Results and discussion: PMB adsorption significantly enhanced the bacteriostatic and bactericidal activity of GO against both planktonic cells and bacterial cells in biofilms. Furthermore, the coatings of PMB-adsorbed GO applied to catheter tubes strongly mitigated biofilm formation, by preventing bacterial adhesion and killing the bacterial cells that managed to attach. The presented results suggest that antibacterial peptide absorption can significantly enhance the antibacterial activity of GO and the resulting material can be effectively used not only against planktonic bacteria but also against infectious biofilms.

Automated Prediction of Bacterial Exclusion Areas on SEM Images of Graphene-Polymer Composites.

To counter the rising threat of bacterial infections in the post-antibiotic age, intensive efforts are invested in engineering new materials with antibacterial properties. The key bottleneck in this initiative is the speed of evaluation of the antibacterial potential of new materials. To overcome this, we developed an automated pipeline for the prediction of antibacterial potential based on scanning electron microscopy images of engineered surfaces. We developed polymer composites containing graphite-oriented nanoplatelets (GNPs). The key property that the algorithm needs to consider is the density of sharp exposed edges of GNPs that kill bacteria on contact. The surface area of these sharp exposed edges of GNPs, accessible to bacteria, needs to be inferior to the diameter of a typical bacterial cell. To test this assumption, we prepared several composites with variable distribution of exposed edges of GNP. For each of them, the percentage of bacterial exclusion area was predicted by our algorithm and validated experimentally by measuring the loss of viability of the opportunistic pathogen Staphylococcus epidermidis. We observed a remarkable linear correlation between predicted bacterial exclusion area and measured loss of viability (R2 = 0.95). The algorithm parameters we used are not generally applicable to any antibacterial surface. For each surface, key mechanistic parameters must be defined for successful prediction.

Silver Nanoparticles: Bactericidal and Mechanistic Approach against Drug Resistant Pathogens.

This review highlights the different modes of synthesizing silver nanoparticles (AgNPs) from their elemental state to particle format and their mechanism of action against multidrug-resistant and biofilm-forming bacterial pathogens. Various studies have demonstrated that the AgNPs cause oxidative stress, protein dysfunction, membrane disruption, and DNA damage in bacteria, ultimately leading to bacterial death. AgNPs have also been found to alter the adhesion of bacterial cells to prevent biofilm formation. The benefits of using AgNPs in medicine are, to some extent, counter-weighted by their toxic effect on humans and the environment. In this review, we have compiled recent studies demonstrating the antibacterial activity of AgNPs, and we are discussing the known mechanisms of action of AgNPs against bacterial pathogens. Ongoing clinical trials involving AgNPs are briefly presented. A particular focus is placed on the mechanism of interaction of AgNPs with bacterial biofilms, which are a significant pathogenicity determinant. A brief overview of the use of AgNPs in other medical applications (e.g., diagnostics, promotion of wound healing) and the non-medical sectors is presented. Finally, current drawbacks and limitations of AgNPs use in medicine are discussed, and perspectives for the improved future use of functionalized AgNPs in medical applications are presented.

Ginsenoside Rg3 Reduces the Toxicity of Graphene Oxide Used for pH-Responsive Delivery of Doxorubicin to Liver and Breast Cancer Cells.

Doxorubicin (DOX) is extensively used in chemotherapy, but it has serious side effects and is inefficient against some cancers, e.g., hepatocarcinoma. To ameliorate the delivery of DOX and reduce its side effects, we designed a pH-responsive delivery system based on graphene oxide (GO) that is capable of a targeted drug release in the acidic tumor microenvironment. GO itself disrupted glutathione biosynthesis and induced reactive oxygen species (ROS) accumulation in human cells. It induced IL17-directed JAK-STAT signaling and VEGF gene expression, leading to increased cell proliferation as an unwanted effect. To counter this, GO was conjugated with the antioxidant, ginsenoside Rg3, prior to loading with DOX. The conjugation of Rg3 to GO significantly reduced the toxicity of the GO carrier by abolishing ROS production. Furthermore, treatment of cells with GO-Rg3 did not induce IL17-directed JAK-STAT signaling and VEGF gene expression-nor cell proliferation-suggesting GO-Rg3 as a promising drug carrier. The anticancer activity of GO-Rg3-DOX conjugates was investigated against Huh7 hepatocarcinoma and MDA-MB-231 breast cancer cells. GO-Rg3-DOX conjugates significantly reduced cancer cell viability, primarily via downregulation of transcription regulatory genes and upregulation of apoptosis genes. GO-Rg3 is an effective, biocompatible, and pH responsive DOX carrier with potential to improve chemotherapy-at least against liver and breast cancers.

Inhibition of bacterial adhesion by epigallocatechin gallate attached polymeric membranes.

Microbial adhesion and formation of biofilms cause a serious problem in several areas including but not limited to food spoilage, industrial corrosion and nosocomial infections. These microbial biofilms pose a serious threat to human health since microbial communities in the biofilm matrix are protected with exopolymeric substances and difficult to eradicate with antibiotics. Hence, the prevention of microbial adhesion followed by biofilm formation is one of the promising strategies to prevent these consequences. The attachment of antimicrobial agents, coatings of nanomaterials and synthesis of hybrid materials are widely used approach to develop surfaces having potential to hinder bacterial adhesion and biofilm formation. In this study, epigallocatechin gallate (EGCG) is attached on p(HEMA-co-GMA) membranes to prevent the bacterial colonization. The attachment of EGCG to membranes was proved by Fourier-transform infrared spectroscopy (FT-IR). The synthesized membrane showed porous structure (SEM), and desirable swelling degree, which are ideal when it comes to the application in biotechnology and biomedicine. Furthermore, EGCG attached membrane showed significant potential to prevent the microbial colonization on the surface. The obtained results suggest that EGCG attached polymer could be used as an alternative approach to prevent the microbial colonization on the biomedical surfaces, food processing equipment as well as development of microbial resistant food packaging systems.

A Road Map toward Field-Effect Transistor Biosensor Technology for Early Stage Cancer Detection.

Field effect transistor (FET)-based nanoelectronic biosensor devices provide a viable route for specific and sensitive detection of cancer biomarkers, which can be used for early stage cancer detection, monitoring the progress of the disease, and evaluating the effectiveness of therapies. On the road to implementation of FET-based devices in cancer diagnostics, several key issues need to be addressed: sensitivity, selectivity, operational conditions, anti-interference, reusability, reproducibility, disposability, large-scale production, and economic viability. To address these well-known issues, significant research efforts have been made recently. An overview of these efforts is provided here, highlighting the approaches and strategies presently engaged at each developmental stage, from the design and fabrication of devices to performance evaluation and data analysis. Specifically, this review discusses the multistep fabrication of FETs, choice of bioreceptors for relevant biomarkers, operational conditions, measurement configuration, and outlines strategies to improve the sensing performance and reach the level required for clinical applications. Finally, this review outlines the expected progress to the future generation of FET-based diagnostic devices and discusses their potential for detection of cancer biomarkers as well as biomarkers of other noncommunicable and communicable diseases.

Cellular and subcellular interactions of graphene-based materials with cancerous and non-cancerous cells.

Despite significant advances in early detection and personalized treatment, cancer is still among the leading causes of death globally. One of the possible anticancer approaches that is presently receiving a lot of attention is the development of nanocarriers capable of specific and efficient delivery of anticancer drugs. Graphene-based materials are promising nanocarriers in this respect, due to their high drug loading capacity and biocompatibility. In this review, we present an overview on the interactions of graphene-based materials with normal mammalian cells at the molecular level as well as cellular and subcellular levels, including plasma membrane, cytoskeleton, and membrane-bound organelles such as lysosomes, mitochondria, nucleus, endoplasmic reticulum, and peroxisome. In parallel, we assemble the knowledge about the interactions of graphene-based materials with cancerous cells, that are considered as the potential applications of these materials for cancer therapy including metastasis treatment, targeted drug delivery, and differentiation to non-cancer stem cells. We highlight the influence of key parameters, such as the size and surface chemistry of graphene-based materials that govern the efficiency of internalization and biocompatibility of these particles in vitro and in vivo. Finally, this review aims to correlate the key parameters of graphene-based nanomaterials specially graphene oxide, such as size and surface modifications, to their interactions with the cancerous and non-cancerous cells for designing and engineering them for bio-applications and especially for therapeutic purposes.

Enriched microbial communities for ammonium and nitrite removal from recirculating aquaculture systems.

The aim of this study was the enrichment of high-performance microbial communities in biofilters for removal of ammonium and nitrite from aquaculture water. Ammonium oxidizing bacteria (AOB) and nitrite oxidizing bacteria (NOB) were enriched from different environmental water samples. The microbial communities with higher ammonium and nitrite removal activity were selected and adapted to different temperatures [9 °C, 15 °C, room temperature (25 °C), and 30 °C]. The expression of genes involved in nitrification including ammonia monooxygenase (AMO) and nitrite oxidoreductase (NXR) were measured in temperature-adapted AOB and NOB microbiomes. The microbial species present in the selected microbiomes were identified via 16s rRNA sequencing. The microbial communities containing Nitrosomonas oligotropha and Nitrobacter winogradskyi showed the highest ammonium and nitrite removal activity at all temperatures used for adaptation. Furthermore, the microbial communities do not contain any pathogenic bacteria. They also exhibited the highest expression of AMO and NXR genes. Using the enriched microbial communities, we achieved a 288% and 181% improvement in ammonium and nitrite removal over the commonly used communities in biofilters at 9 °C, respectively. These results suggest that the selected microbiomes allowed for a significant improvement of water quality in a recirculating aquaculture system (RAS).

Insights into the Mechanism for Vertical Graphene Growth by Plasma-Enhanced Chemical Vapor Deposition.

Vertically oriented graphene (VG) has attracted attention for years, but the growth mechanism is still not fully revealed. The electric field may play a role, but the direct evidence and exactly what role it plays remains unclear. Here, we conduct a systematic study and find that in plasma-enhanced chemical vapor deposition, the VG growth preferably occurs at spots where the local field is stronger, for example, at GaN nanowire tips. On almost round-shaped nanoparticles, instead of being perpendicular to the substrate, the VG grows along the field direction, that is, perpendicular to the particles' local surfaces. Even more convincingly, the sheath field is screened to different degrees, and a direct correlation between the field strength and the VG growth is observed. Numerical calculation suggests that during the growth, the field helps accumulate charges on graphene, which eventually changes the cohesive graphene layers into separate three-dimensional VG flakes. Furthermore, the field helps attract charged precursors to places sticking out from the substrate and makes them even sharper and turn into VG. Finally, we demonstrate that the VG-covered nanoparticles are benign to human blood leukocytes and could be considered for drug delivery. Our research may serve as a starting point for further vertical two-dimensional material growth mechanism studies.

Antimicrobial Activity of Graphene Oxide Contributes to Alteration of Key Stress-Related and Membrane Bound Proteins.

Introduction: Antibacterial activity of graphene oxide (GO) has been extensively studied, wherein penetration of the bacterial cell membrane and oxidative stress are considered to play a major role in the bactericidal activity of GO. However, the specific mechanism responsible for the antibacterial activity of GO remains largely unknown. Hence, the goal of this study was to explore the mode of action of GO, via an in-depth proteomic analysis of the targeted bacteria. Methods: Staphylococcus aureus was grown in the presence of GO and samples were collected at different growth phases to examine the cell viability and to analyze the changes in protein expression. Antimicrobial efficiency of GO was tested by assessing bacterial viability, live/dead staining and scanning electron microscopy. The intracellular reactive oxygen species (ROS) induced by GO treatment were examined by fluorescence microscopy. Label-free quantitative proteomics analysis was performed to examine the differentially regulated proteins in S. aureus after GO treatment. Results: GO treatment was observed to reduce S. aureus viability, from 50 ± 17% after 4 h, to 93 ± 2% after 24 h. The live/dead staining confirmed this progressive antimicrobial effect of GO. SEM images revealed the wrapping of bacterial cells and their morphological disruption by means of pore formation due to GO insertion. GO treatment was observed to generate intracellular ROS, correlating to the loss of cell viability. The proteomics analysis revealed alteration in the expression of cell membrane, oxidative stress response, general stress response, and virulence-associated proteins in GO-treated bacterial cells. The time-dependent bactericidal activity of GO correlated with a higher number of differentially regulated proteins involved in the above.-mentioned processes. Conclusion: The obtained results suggest that the time-dependent bactericidal effect of GO is attributed to its wrapping/trapping ability, ROS production and due to physical disruption of the cell membrane.

Graphene-Based Sensor for Detection of Bacterial Pathogens.

Microbial colonization to biomedical surfaces and biofilm formation is one of the key challenges in the medical field. Recalcitrant biofilms on such surfaces cause serious infections which are difficult to treat using antimicrobial agents, due to their complex structure. Early detection of microbial colonization and monitoring of biofilm growth could turn the tide by providing timely guidance for treatment or replacement of biomedical devices. Hence, there is a need for sensors, which could generate rapid signals upon bacterial colonization. In this study, we developed a simple prototype sensor based on pristine, non-functionalized graphene. The detection principle is a change in electrical resistance of graphene upon exposure to bacterial cells. Without functionalization with specific receptors, such sensors cannot be expected to be selective to certain bacteria. However, we demonstrated that two different bacterial species can be detected and differentiated by our sensor due to their different growth dynamics, adherence pattern, density of adhered bacteria and microcolonies formation. These distinct behaviors of tested bacteria depicted distinguishable pattern of resistance change, resistance versus gate voltage plot and hysteresis effect. This sensor is simple to fabricate, can easily be miniaturized, and can be effective in cases when precise identification of species is not needed.

Graphene coated magnetic nanoparticles facilitate the release of biofuels and oleochemicals from yeast cell factories.

Engineering of microbial cells to produce high value chemicals is rapidly advancing. Yeast, bacteria and microalgae are being used to produce high value chemicals by utilizing widely available carbon sources. However, current extraction processes of many high value products from these cells are time- and labor-consuming and require toxic chemicals. This makes the extraction processes detrimental to the environment and not economically feasible. Hence, there is a demand for the development of simple, effective, and environmentally friendly method for the extraction of high value chemicals from these cell factories. Herein, we hypothesized that atomically thin edges of graphene having ability to interact with hydrophobic materials, could be used to extract high value lipids from cell factories. To achieve this, array of axially oriented graphene was deposited on iron nanoparticles. These coated nanoparticles were used to facilitate the release of intracellular lipids from Yarrowia lipolytica cells. Our treatment process can be integrated with the growth procedure and achieved the release of 50% of total cellular lipids from Y. lipolytica cells. Based on this result, we propose that nanoparticles coated with axially oriented graphene could pave efficient, environmentally friendly, and cost-effective way to release intracellular lipids from yeast cell factories.

Silver nanoparticles produced from Cedecea sp. exhibit antibiofilm activity and remarkable stability.

With multidrug-resistant bacterial pathogens on the rise, there is a strong research focus on alternative antibacterial treatments that could replace or complement classical antibiotics. Metallic nanoparticles, and in particular silver nanoparticles (AgNPs), have been shown to kill bacterial biofilms effectively, but their chemical synthesis often involves environmentally unfriendly by-products. Recent studies have shown that microbial and plant extracts can be used for the environmentally friendly synthesis of AgNPs. Herein we report a procedure for producing AgNPs using a putative Cedecea sp. strain isolated from soil. The isolated bacterial strain showed a remarkable potential for producing spherical, crystalline and stable AgNPs characterized by UV-visible spectroscopy, transmission electron microscopy, dynamic light scattering, and Fourier transform infrared spectroscopy. The concentration of produced nanoparticles was 1.31 µg/µl with a negative surface charge of - 15.3 mV and nanoparticles size ranging from 10-40 nm. The AgNPs was tested against four pathogenic microorganisms S. epidermidis, S. aureus, E. coli and P. aeruginosa. The nanoparticles exhibited strong minimum inhibitory concentration (MIC) values of 12.5 and 6.25 µg/µl and minimum bactericidal concentration (MBC) values of 12.5 and 12.5 µg/mL against E. coli and P. aeruginosa, respectively. One distinguishing feature of AgNPs produced by Cedecea sp. extracts is their extreme stability. Inductively coupled plasma mass spectrometry and thermogravimetric analysis demonstrated that the produced AgNPs are stable for periods exceeding one year. This means that their strong antibacterial effects, demonstrated against E. coli and P. aeruginosa biofilms, can be expected to persist during extended periods.

Sustained release of usnic acid from graphene coatings ensures long term antibiofilm protection.

Protecting surfaces from bacterial colonization and biofilm development is an important challenge for the medical sector, particularly when it comes to biomedical devices and implants that spend longer periods in contact with the human body. A particularly difficult challenge is ensuring long-term protection, which is usually attempted by ensuring sustained release of antibacterial compounds loaded onto various coatings. Graphene have a considerable potential to reversibly interact water insoluble molecules, which makes them promising cargo systems for sustained release of such compounds. In this study, we developed graphene coatings that act as carriers capable of sustained release of usnic acid (UA), and hence enable long-term protection of surfaces against colonization by bacterial pathogens Staphylococcus aureus and Staphylococcus epidermidis. Our coatings exhibited several features that made them particularly effective for antibiofilm protection: (i) UA was successfully integrated with the graphene material, (ii) a steady release of UA was documented, (iii) steady UA release ensured strong inhibition of bacterial biofilm formation. Interestingly, even after the initial burst release of UA, the second phase of steady release was sufficient to block bacterial colonization. Based on these results, we propose that graphene coatings loaded with UA can serve as effective antibiofilm protection of biomedical surfaces.